Characterization of purified cathepsin D from malignant human breast tissue.

  • Authors:
    • S Bazel
    • J A Alhadeff
  • View Affiliations

  • Published online on: February 1, 1999     https://doi.org/10.3892/ijo.14.2.315
  • Pages: 315-324
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Abstract

The aspartyl protease cathepsin D (EC 3.4.23.5) appears to be found in increased amounts and/or abnormally secreted in breast cancer cells, and may contribute to the metastatic spread of malignancy. In the present study, cathepsin D was purified 4800-fold in 20% yield from malignant human breast tissue using affinity chromatography on pepstatin-agarose and DEAE-Sephadex chromatography. Slab gel SDS-PAGE of the purified cathepsin D indicated the presence of three major protein bands (31, 13, 12, kDa) and two minor protein bands (47, 29 kDa). Western blotting indicated that the 31 kDa band was the major immunoreactive species. Isoelectric focusing indicated that the purified cathepsin D consisted of three major isoforms at approximate pIs of 7.4, 7.0 and 6.6, and a possible isoform of lower activity centered around pI 3.2. The pH curve of purified cathepsin D indicated a broad optimum centered around pH 3.4. Lectin blotting suggested the presence of mannose residues but no evidence was found for lectin-available sialic acid, fucose, N-acetylglucosamine and galactose residues. The investigated properties of purified cathepsin D from malignant breast tissue are very similar, if not identical, to the properties of cathepsin D previously purified from normal human breast tissue. Our findings suggest that the elevated activity and antigenic levels of cathepsin D in malignant breast tissue are due to increased amounts of apparently normal enzyme.

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February 1999
Volume 14 Issue 2

Print ISSN: 1019-6439
Online ISSN:1791-2423

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APA
Bazel, S., & Bazel, S. (1999). Characterization of purified cathepsin D from malignant human breast tissue.. International Journal of Oncology, 14, 315-324. https://doi.org/10.3892/ijo.14.2.315
MLA
Bazel, S., Alhadeff, J."Characterization of purified cathepsin D from malignant human breast tissue.". International Journal of Oncology 14.2 (1999): 315-324.
Chicago
Bazel, S., Alhadeff, J."Characterization of purified cathepsin D from malignant human breast tissue.". International Journal of Oncology 14, no. 2 (1999): 315-324. https://doi.org/10.3892/ijo.14.2.315